Development and characterization of SV40 immortalized rat submandibular acinar cell lines

Quissell, David O.; Barzen, Katherine A.; Gruenert, Dieter C.; Redman, Robert S.; Camden, Jean M.; Turner, John T.

doi:10.1007/s11626-997-0137-8

Cited by 77 publications

(89 citation statements)

References 18 publications

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“…Because there is no appropriate cell line derived from either human or rabbit SMGs for further study on tight-junction structure and function, we chose the rat SMG-C6 cells to characterize the effect of TRPV1 on SMG epithelia. This cell line, originally established by Quissel et al, was reported to maintain a normal acinar function and have similar characteristics, including tight junction structure, as determined by electron microscopy (Quissell et al, 1997). In the present study, we confirmed the expression of TRPV1 and occludin and characterized their distribution.…”

Section: Discussionsupporting

confidence: 83%

Occludin is required for TRPV1-modulated paracellular permeability in the submandibular gland

Cong

Zhang

Yang

et al. 2013

Journal of Cell Science

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SummaryOccludin plays an important role in maintaining tight junction barrier function in many types of epithelia. We previously reported that activation of transient receptor potential vanilloid subtype 1 (TRPV1) in rabbit submandibular gland promoted salivary secretion, partly by an increase in paracellular permeability. We have now explored the role of occludin in TRPV1-modulated paracellular permeability in a rat submandibular gland cell line SMG-C6. Both TRPV1 and occludin were expressed in SMG-C6 cells, and capsaicin induced redistribution of occludin, but not claudin-3, claudin-4 or E-cadherin, from the cell membrane into the cytoplasm. Capsaicin also decreased transepithelial electrical resistance (TER) and increased the Trypan Blue and FITC-dextran flux. Capsazepine (CPZ), a TRPV1 antagonist, inhibited the capsaicin-induced occludin redistribution and TER decrease. Moreover, occludin knockdown by shRNA suppressed, whereas occludin re-expression restored, the TER response to capsaicin. Mechanistically, TRPV1 activation increased ERK1/2 and MLC2 phosphorylation. PD98059, an ERK1/2 kinase inhibitor, abolished the capsaicin-induced MLC2 phosphorylation, whereas ML-7, an MLC2 kinase inhibitor, did not affect ERK1/2 phosphorylation, suggesting that ERK1/2 is the upstream signaling molecule of MLC2. Capsaicin also induced F-actin reorganization, which was abolished by CPZ, PD98059 and ML-7, indicating that TRPV1 activation altered F-actin organization in an ERK1/2-and MLC2-dependent manner. Furthermore, either PD98059 or ML-7 could abolish the capsaicin-induced TER response and occludin redistribution, whereas knockdown of ERK1/2 further confirmed that the TRPV1-modulated paracellular permeability was ERK1/2 dependent. Taken together, these results identified a crucial role of occludin in submandibular epithelial cells, and more importantly, demonstrated that occludin was required to mediate TRPV1-modulated paracellular permeability.

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Section: Discussionsupporting

confidence: 83%

Occludin is required for TRPV1-modulated paracellular permeability in the submandibular gland

Cong

Zhang

Yang

et al. 2013

Journal of Cell Science

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“…), and VOL. 26,2006 SUPPRESSION OF SALIVARY ACINAR CELL APOPTOSIS BY Akt 8841 primary parotid or submandibular acinar cells were prepared under sterile conditions similar to previously published protocols (50,51,72 (3,51). Activation of caspase 3 was quantitated with the BioMol QuantiZyme Colorimetric assay kit (Plymouth Meeting, PA).…”

Section: Methodsmentioning

confidence: 99%

MDM2 Is Required for Suppression of Apoptosis by Activated Akt1 in Salivary Acinar Cells

Limesand¹,

Schwertfeger

Anderson

2006

Molecular and Cellular Biology

111

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Chronic damage to the salivary glands is a common side effect following head and neck irradiation. It is hypothesized that irreversible damage to the salivary glands occurs immediately after radiation; however, previous studies with rat models have not shown a causal role for apoptosis in radiation-induced injury. We report that etoposide and gamma irradiation induce apoptosis of salivary acinar cells from FVB control mice in vitro and in vivo; however, apoptosis is reduced in transgenic mice expressing a constitutively activated mutant of Akt1 (myr-Akt1). Expression of myr-Akt1 in the salivary glands results in a significant reduction in phosphorylation of p53 at serine 18 , total p53 protein accumulation, and p21 WAF1 or Bax mRNA following etoposide or gamma irradiation of primary salivary acinar cells. The reduced level of p53 protein in myr-Akt1 salivary glands corresponds with an increase in MDM2 phosphorylation in vivo, suggesting that the Akt/ MDM2/p53 pathway is responsible for suppression of apoptosis. Dominant-negative Akt blocked phosphorylation of MDM2 in salivary acinar cells from myr-Akt1 transgenic mice. Reduction of MDM2 levels in myr-Akt1 primary salivary acinar cells with small interfering RNA increases the levels of p53 protein and renders these cells susceptible to etoposide-induced apoptosis in spite of the presence of activated Akt1. These results indicate that MDM2 is a critical substrate of activated Akt1 in the suppression of p53-dependent apoptosis in vivo.

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“…Mice were anesthetized with sodium pentobarbital (60 mg/kg) and primary parotid acinar cells were prepared under sterile conditions similar to previously published protocols used in the preparation of primary acinar cells from rat salivary glands. 48 A 1% vol/vol cell suspension was seeded onto collagen-coated dishes (Falcon/Becton Dickenson) in media similar to that used for the established cell lines with the exception of the FBS content elevated from 2.5 to 10%. Cells reached approximately 80% confluency after 5 days in culture and were utilized immediately in experiments at that time without passage.…”

Section: Cells and Cell Culturementioning

confidence: 99%

Synergistic suppression of apoptosis in salivary acinar cells by IGF1 and EGF

et al. 2003

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Tissue homeostasis requires balancing cell proliferation and programmed cell death. IGF1 significantly suppressed etoposide-induced apoptosis, measured by caspase 3 activation and quantitation of cellular subG 1 DNA content, in rat parotid salivary acinar cells (C5). Transduction of C5 cells with an adenovirus expressing a constitutively activated mutant of Akt-suppressed etoposide-induced apoptosis, whereas a kinase-inactive mutant of Akt suppressed the protective effect of IGF1. IGF1 also suppressed apoptosis induced by taxol and brefeldin A. EGF was unable to suppress apoptosis induced by etoposide, but was able to synergize with IGF1 to further suppress caspase 3 activation and DNA cleavage after etoposide treatment. The catalytic activity of Akt was significantly higher following stimulation with both growth factors compared to stimulation with IGF1 or EGF alone. These results suggest that a threshold of activated Akt is required for suppression of apoptosis and the cooperative action of growth factors in regulating salivary gland homeostasis.

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Development and characterization of SV40 immortalized rat submandibular acinar cell lines

Cited by 77 publications

References 18 publications

Occludin is required for TRPV1-modulated paracellular permeability in the submandibular gland

Occludin is required for TRPV1-modulated paracellular permeability in the submandibular gland

MDM2 Is Required for Suppression of Apoptosis by Activated Akt1 in Salivary Acinar Cells

Synergistic suppression of apoptosis in salivary acinar cells by IGF1 and EGF

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